The platynosomiasis, a worldwide parasitic disease with importance for domestic cat, has an etiological agent species of trematodes of the genus Platynosomum, whose complete life cycles are not yet known. The real role of lizards in the transmission of this dicrocoeliid parasite (as obligatory intermediate or paratenic host) still needs to be defined. In the present study, oval-shaped encysted metacercariae obtained from terrestrial isopods (Oniscidea sp. and Nagurus nanus) and elongated excysted metacercariae found in biliary ducts and gallbladder of lizards (Hemidactylus mabouia) in Brazil were used for morphological characterization and experimental infection of mice. Adult parasites recovered from bile ducts and liver of mice inoculated orally with metacercariae from both hosts (isopods and lizards) were identified as Platynosomum illiciens (=Platynosomum fastosum), showing that lizards are paratenic (not obligatory) hosts involved in the life cycle of this parasite. Moreover, Subulina octona is reported as the first intermediate host of P. illiciens in South America, and terrestrial isopods are presented here as new natural second intermediate hosts of the parasite. Finally, it is pointed out that high prevalence and intensity of infection of intermediate and paratenic hosts were observed. These findings on the life cycle of P. illiciens are relevant considering that they may indicate possible control measures of platynosomiasis.